转染的FasL基因诱导HCC9204细胞凋亡的形态学研究

目的 构建含FasL基因的腺病毒载体，转染肝癌9204细胞株，观察FasL基因对9204细胞株的影响。方法 以腺病毒作载体，把FasL基因克隆入腺病毒载体，构建成含FasL基因的转基因载体并酶切鉴定，用脂质体包裹法把含FasL的腺病毒载体转染HCC9204细胞株，用RT－PCR方法证实转染后有无FasL基因表达，采用电镜观察9204细胞形态学变化。结果 酶切鉴定证实含FasL基因的转基因载体构建成功；RT－PCR显示转染后9204细胞株FasL基因表达水平显高于未转染细胞株（P＜0.01）；肝癌细胞株9204转染FasL基因72h后，在光镜下可见调亡细胞体积缩小，生长不良，染色质浓度，密度增高，胞核中出现颗粒样物质，电镜下可见调亡小体。结论 成功地构建了FasL基因的转基因载体；转染FasL基因可诱导肝癌9204细胞株出现凋亡。

Morphological studies on apoptosis of HCC8204 cell line induced by transferring FasL into HCC9204 cell line

AIM To construct adenovirus vector with FasL, transfer HCC9204 cell line, and observe FasL effects on HCC9204. METHODS Using adenovirus as vector, we cloned FasL gene into vector and identified if FasL gene was cloned into vector by using enzyme; we used Lipofectin to pack vector with FasL, transferred vector into HCC9204 cell line, and then observed morphologic change of HCC9204 cell. RESULTS Enzyme digestion results showed that FasL gene was cloned into adenovirus vector; 72 hours after transferring FasL gene into HCC9204 cell line,FasL gene was expressed at a higher level than that in untransferred cells ( P <0.01); condensation of chromosome and appotosis body were observed under electroscope. CONCLUSION Adenovirus vector with FasL gene is successfully constructed; transferred FasL may induce HCC9204 cell line to display cell appotosis.